LOCUS       sequence-375354-        4850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Clostridium expression vector (pCB102 origin, specR) with drop-out 
            Plac-RFP cassette flanked with BsaI sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4850)
  AUTHORS   Rostain W, Zaplana T, Boutard M, Baum C, Tabuteau S, Sanitha M, 
            Ramya M, Guss A, Ettwiller L, Tolonen AC
  TITLE     Tuning of Gene Expression in Clostridium phytofermentans Using 
            Synthetic Promoters and CRISPRi.
  JOURNAL   ACS Synth Biol. 2022 Dec 16;11(12):4077-4088. doi: 
            10.1021/acssynbio.2c00385. Epub 2022 Nov 25.
  PUBMED    36427328
REFERENCE   2  (bases 1 to 4850)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.2c00385"; journalName: "ACS Synth Biol"; date: 
            "2022-12-16- 16"; volume: "11"; issue: "12"; pages: "4077-4088"
COMMENT     Sequence Label: pQmod3S-GG
FEATURES             Location/Qualifiers
     source          1..4850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    236..257
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(272..277,278..295,296..302)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    310..326
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             344..355
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             362..1039
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1084..1155
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1171..1198
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      4060..4648
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4549..4568
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4802..4819
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cctgcaggat aaaaaaattg tagataaatt ttataaaata gttttatcta caattttttt
       61 atcaggaaac agctatgacc gcggccgctg tatccatatg accatgatta cgaattcgag
      121 ctcaggatga gacctcaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      181 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      241 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
      301 tgtgtggaat tgtgagcgga taacaatttc acacatacta gagaaagagg agaaatacta
      361 gatggcttcc tccgaagacg ttatcaaaga gttcatgcgt ttcaaagttc gtatggaagg
      421 ttccgttaac ggtcacgagt tcgaaatcga aggtgaaggt gaaggtcgtc cgtacgaagg
      481 tacccagacc gctaaactga aagttaccaa aggtggtccg ctgccgttcg cttgggacat
      541 cctgtccccg cagttccagt acggttccaa agcttacgtt aaacacccgg ctgacatccc
      601 ggactacctg aaactgtcct tcccggaagg tttcaaatgg gaacgtgtta tgaacttcga
      661 agacggtggt gttgttaccg ttacccagga ctcctccctg caagacggtg agttcatcta
      721 caaagttaaa ctgcgtggta ccaacttccc gtccgacggt ccggttatgc agaaaaaaac
      781 catgggttgg gaagcttcca ccgaacgtat gtacccggaa gacggtgctc tgaaaggtga
      841 aatcaaaatg cgtctgaaac tgaaagacgg tggtcactac gacgctgaag ttaaaaccac
      901 ctacatggct aaaaaaccgg ttcagctgcc gggtgcttac aaaaccgaca tcaaactgga
      961 catcacctcc cacaacgaag actacaccat cgttgaacag tacgaacgtg ctgaaggtcg
     1021 tcactccacc ggtgcttaat aacgctgata gtgctagtgt agatcgctac tagagccagg
     1081 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     1141 tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt
     1201 tataactatg gtctcccgaa tcttccgcta gcataaaaat aagaagcctg catttgcagg
     1261 cttcttattt ttatggcgcg ccgccattat ttttttgaac aattgacaat tcatttctta
     1321 ttttttatta agtgatagtc aaaaggcata acagtgctga atagaaagaa atttacagaa
     1381 aagaaaatta tagaatttag tatgattaat tatactcatt tatgaatgtt taattgaata
     1441 caaaaaaaaa tacttgttat gtattcaatt acgggttaaa atatagacaa gttgaaaaat
     1501 ttaataaaaa aataagtcct cagctcttat atattaagct accaacttag tatataagcc
     1561 aaaacttaaa tgtgctacca acacatcaag ccgttagaga actctatcta tagcaatatt
     1621 tcaaatgtac cgacatacaa gagaaacatt aactatatat attcaattta tgagattatc
     1681 ttaacagata taaatgtaaa ttgcaataag taagatttag aagtttatag cctttgtgta
     1741 ttggaagcag tacgcaaagg cttttttatt tgataaaaat tagaagtata tttatttttt
     1801 cataattaat ttatgaaaat gaaagggggt gagcaaagtg acagaggaaa gcagtatctt
     1861 atcaaataac aaggtattag caatatcatt attgacttta gcagtaaaca ttatgacttt
     1921 tatagtgctt gtagctaagt agtacgaaag ggggagcttt aaaaagctcc ttggaataca
     1981 tagaattcat aaattaattt atgaaaagaa gggcgtatat gaaaacttgt aaaaattgca
     2041 aagagtttat taaagatact gaaatatgca aaatacattc gttgatgatt catgataaaa
     2101 cagtagcaac ctattgcagt aaatacaatg agtcaagatg tttacataaa gggaaagtcc
     2161 aatgtattaa ttgttcaaag atgaaccgat atggatggtg tgccataaaa atgagatgtt
     2221 ttacagagga agaacagaaa aaagaacgta catgcattaa atattatgca aggagcttta
     2281 aaaaagctca tgtaaagaag agtaaaaaga aaaaataatt tatttattaa tttaatattg
     2341 agagtgccga cacagtatgc actaaaaaat atatctgtgg tgtagtgagc cgatacaaaa
     2401 ggatagtcac tcgcattttc ataatacatc ttatgttatg attatgtgtc ggtgggactt
     2461 cacgacgaaa acccacaata aaaaaagagt tcggggtagg gttaagcata gttgaggcaa
     2521 ctaaacaatc aagctaggat atgcagtagc agaccgtaag gtcgttgttt aggtgtgttg
     2581 taatacatac gctattaaga tgtaaaaata cggataccaa tgaagggaaa agtataattt
     2641 ttggatgtag tttgtttgtt catctatggg caaactacgt ccaaagccgt ttccaaatct
     2701 gctaaaaagt atatcctttc taaaatcaaa gtcaagtatg aaatcataaa taaagtttaa
     2761 ttttgaagtt attatgatat tatgtttttc tattaaaata aattaagtat atagaatagt
     2821 ttaataatag tatatactta atgtgataag tgtctgacag tgtcacagaa aggatgattg
     2881 ttatggatta taagcggccg gcccaatgaa taggtttaca cttactttag ttttatggaa
     2941 atgaaagatc atatcatata taatctagaa taaaattaac taaataatta ttatctagat
     3001 aaaaaattta gaagccaatg aaatctataa ataaactaaa ttaagtttat ttaattaaca
     3061 actatggata taaaataggt actaatcaaa atagtgagga ggatatattt gaatacatac
     3121 gaacaaatta ataaagtgaa aaaaatactt cggaaacatt taaaaaataa ccttattggt
     3181 acttacatgt ttggatcagg agttgagagt ggactaaaac caaatagtga tcttgacttt
     3241 ttagtcgtcg tatctgaacc attgacagat caaagtaaag aaatacttat acaaaaaatt
     3301 agacctattt caaaaaaaat aggagataaa agcaacttac gatatattga attaacaatt
     3361 attattcagc aagaaatggt accgtggaat catcctccca aacaagaatt tatttatgga
     3421 gaatggttac aagagcttta tgaacaagga tacattcctc agaaggaatt aaattcagat
     3481 ttaaccataa tgctttacca agcaaaacga aaaaataaaa gaatatacgg aaattatgac
     3541 ttagaggaat tactacctga tattccattt tctgatgtga gaagagccat tatggattcg
     3601 tcagaggaat taatagataa ttatcaggat gatgaaacca actctatatt aactttatgc
     3661 cgtatgattt taactatgga cacgggtaaa atcataccaa aagatattgc gggaaatgca
     3721 gtggctgaat cttctccatt agaacatagg gagagaattt tgttagcagt tcgtagttat
     3781 cttggagaga atattgaatg gactaatgaa aatgtaaatt taactataaa ctatttaaat
     3841 aacagattaa aaaaattata aaaaaattga aaaaatggtg gaaacacttt tttcaatttt
     3901 tttgttttat tatttaatat ttgggaaata ttcattctaa ttggtaatca gattttagaa
     3961 gtttaaactc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4021 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4081 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4141 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4201 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4261 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4321 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4381 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4441 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4501 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4561 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4621 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4681 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4741 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4801 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcagggccc
//
